College of Animal Science and Technology, Zhejiang Agriculture and Forestry University, Hangzhou 311300, China.
Sci Rep. 2017 Feb 9;7:42334. doi: 10.1038/srep42334.
In this study, we discovered that shrimp miR-100 was up-regulated at 24 h after WSSV or Vibrio alginolyticus infection, confirming its participation in the innate immune system of shrimp. The anti-miRNA oligonucleotide (AMO-miR-100) was applied to inhibit the expression of miR-100. After AMO-miR-100 treatment, the shrimp was challenged with WSSV or V. alginolyticus. The knockdown of miR-100 expression decreased the mortality of WSSV-infected shrimp from 24 h to 72 h post-infection and enhanced the mortality of V. alginolyticus-infected shrimp significantly. The knockdown of miR-100 affected phenoloxidase (PO) activity, superoxide dismutase (SOD) activity and total hemocyte count (THC) after the infection with WSSV or V. alginolyticus, indicating a regulative role of miR-100 in the immune potential of shrimp in the response to WSSV or V. alginolyticus infection. The knockdown of miR-100 induced the apoptosis of shrimp hemocytes, and V. alginolyticus + AMO-miR-100 treatment caused more hemocyte apoptosis than V. alginolyticus treatment. The miR-100 influenced also the morphology of shrimp hemocytes and regulated the phagocytosis of WSSV or V. alginolyticus. Thus, we concluded that miR-100 may promote the anti-Vibrio immune response of shrimp through regulating apoptosis, phagocytosis and PO activity and affects the progression of WSSV infection at a certain level.
在这项研究中,我们发现虾 miR-100 在 WSSV 或溶藻弧菌感染后 24 小时上调,证实其参与了虾的先天免疫系统。反义 miRNA 寡核苷酸(AMO-miR-100)被用于抑制 miR-100 的表达。在用 AMO-miR-100 处理后,虾受到 WSSV 或 V. alginolyticus 的挑战。miR-100 表达的敲低降低了 WSSV 感染虾从感染后 24 小时到 72 小时的死亡率,并显著增加了 V. alginolyticus 感染虾的死亡率。miR-100 的敲低影响了感染 WSSV 或 V. alginolyticus 后酚氧化酶(PO)活性、超氧化物歧化酶(SOD)活性和总血细胞计数(THC),表明 miR-100 在虾对 WSSV 或 V. alginolyticus 感染的免疫潜力中具有调节作用。miR-100 的敲低诱导了虾血细胞的凋亡,而 V. alginolyticus+AMO-miR-100 处理比 V. alginolyticus 处理引起更多的血细胞凋亡。miR-100 还影响了虾血细胞的形态,并调节了 WSSV 或 V. alginolyticus 的吞噬作用。因此,我们得出结论,miR-100 可能通过调节凋亡、吞噬作用和 PO 活性来促进虾的抗弧菌免疫反应,并在一定程度上影响 WSSV 感染的进展。